Apparatus and Method for Interleaving Detection of Fluorescence and Luminescence

• US 20080179539A1
• Filed: 01/25/2007
• Published: 07/31/2008
• Est. Priority Date: 01/25/2007
• Status: Active Grant

• Current Assignee: EtaLuma, Inc.
• Original Assignee: Amgen Inc.
• Inventors: Li, Chuck Z., Rasnow, Brian
• Granted Patent: US 7,700,928 B2
• Time in Patent Office: Days
• US Class Current: 250/458.1
• CPC Class Codes: G01N 21/6454 using an integrated detector array

Abstract

An apparatus is provided that is capable of interleaving detection of fluorescence and luminescence signals emitted from a plurality of samples. The apparatus is suitable for analysis of samples containing single cells or tissues up to and including living organisms. It contains an optical assembly or “sandwich” for producing a spectrally pure and spatially disperse light source for illuminating the sample. The invention also provides a plurality of optical sandwiches that can be variously geometrically arranged and their intensities programmed to create spatially uniform illumination over a large sample. The invention further provides an apparatus having at least one of the optical sandwich and a detector system capable of interleaving detection of fluorescent and luminescent signals when a suitable sample is illuminated by the light source of the optical sandwich. Methods for preparing samples and using the sandwiches, arrays and apparatus, are further provided by this invention. A method for interleaving detection of fluorescent and luminescent signals emitted from a plurality of samples is disclosed.

Claims

1. An optical sandwich comprising:

(i) a light source;

(ii) a high numerical aperture collimation lens;

(iii) an excitation filter selected from the group consisting of a sharp spectral band pass, a low pass filter and a high pass filter; and

(iv) a diffuser,

wherein the optical elements (i), (ii), (iii) and (iv) are arranged to provide a spectrally pure and spatially dispersed illumination.

2. The optical sandwich of claim 1, wherein the diffuser is a holographic diffuser.

3. The optical sandwich of claim 1, wherein the light source is selected from the group consisting of an ultra-bright light emitting diode (LED), a deuterium tube, a flash lamp and an ultraviolet LED, a quartz-tungsten halogen (QTH) bulb, a laser diode, and a fluorescent light source.

4. An optical array comprising a plurality of optical sandwiches of claim 1.

5. A plurality of optical arrays of claim 4.

6. The plurality of optical arrays of claim 5, wherein at least two arrays emit light of different wavelength.

7. A rotating shaft comprising a plurality of optical arrays of claim 5.

8. A linear actuator comprising a plurality of optical arrays of claim 5.

9. An apparatus capable of interleaving detection of fluorescence and luminescence emitted from a plurality of samples, the apparatus comprising:

(i) a sample container receiving device;

(ii) an optical sandwich of claim 1; and

(iii) a detector system capable of interleaving detection of fluorescent and luminescent signals emitted from at least one of the plurality of the samples, the detector system comprising;

(i) a detector lens (ii) an emission filter and (iii) a sensor.

10. The apparatus of claim 9, wherein the detector system is an array detector.

11. The apparatus of claim 9, wherein the detector system is an imaging detector.

12. The apparatus of claim 9, wherein the detector system is remotely connected to the optical sandwich so that illumination and detection are synchronized.

13. The apparatus of claim 9, comprising a plurality of optical sandwiches arranged in an array.

14. The apparatus of claim 13, further comprising a plurality of optical arrays.

15. The apparatus of claim 14, wherein the plurality of optical arrays are arranged on a linear actuator or on a rotating shaft.

16. The apparatus of any of claims 13, 14 or 15, wherein at least two of the optical sandwiches are positioned on opposing sides of the sample receiving device.

17. The apparatus of claim 14, wherein at least two of the arrays emit a light source of different wavelengths.

18. The apparatus of claim 9, wherein the emission filter is remotely programmed.

19. The apparatus of claim 9, wherein the emission filter is a rotating emission filter wheel.

20. The apparatus of claim 19, wherein the rotating emission filter wheel is positioned in front of or behind the detector lens.

21. The apparatus of claim 9, further comprising a plurality of emission filters and wherein each filter is positioned on a rotating emission filter wheel.

22. The apparatus of claim 21, wherein the rotating emission filter wheels are coaxial and positioned both in front and behind the detector lens.

23. The apparatus of claim 21, wherein the rotating emission filter wheel is positioned between the sample receiving device and the detector system.

24. The apparatus of claim 21, further comprising at least two rotating emission filter wheels and wherein at least one rotating emission filter wheel is positioned above the detector lens and at least one rotating emission filter wheel is positioned below the detector lens.

25. The apparatus of claim 9, further comprising a feed-back mechanism to control the spatial pattern of illumination of the excitation source from a signal received from the detector system.

26. The apparatus of claim 9, further comprising a sample contained within a sample container.

27. The apparatus of claim 26, wherein the sample comprises a plurality of samples selected from a cell or a plurality of cells.

28. The apparatus of claim 26, wherein the sample container is selected from the group of a slide, a micro titer plate, an agar gel and a cuvette.

29. The apparatus of claim 9, further comprising an apparatus to remove condensation or fog from the sample container.

30. The apparatus of claim 29, wherein the apparatus to remove condensation or fog from the sample container is selected from the group consisting of a wiper mechanism, an evaporative source and a combination thereof.

31. The apparatus of claim 30, wherein the evaporative source comprises a heat source or a blower.

32. The apparatus of claim 9, wherein the detector lens comprises a remotely controlled lens aperture.

33. A method for detecting a signal emitted from a fluorescently labeled sample, comprising illuminating a suitable sample with an excitation source emitted by the light source of the apparatus of claim 9 and detecting any fluorescent signal emitted therefrom.

34. A method for detecting a signal emitted from a luminescently labeled sample, comprising detecting the luminescent signal emitted from a sample comprising a luminescent label with the apparatus of claim 9.

35. A method for detecting a signal emitted from a fluorescently and luminescently labeled sample, comprising illuminating a suitable sample with an excitation source emitted by the light source of the apparatus of claim 9 and detecting a fluorescence or luminescence signal emitted from the sample and wherein the light source is automatically turned off or shuttered during interleaved luminescence imaging.

36. A method comprising interleaving detection of fluorescent and luminescent signals emitted from a plurality of samples, comprising detecting fluorescent and luminescent signals produced by the plurality of samples using the apparatus of claim 9.

37. The method of claim 35 or 36, wherein the sample comprises two different fluorescent labels.

38. The method of claim 37, wherein the plurality of samples express or are labeled with more than one luminescent label.

39. The method of claim 35 or 36, wherein the plurality of samples express or are labeled with one or more fluorescent label and with one or more luminescent label.

40. The method of claim 35, wherein the luminescent label is a bioluminescent label or a chemical luminescent label.

41. The method of claim 36, wherein the luminescent label is a bioluminescent label or a chemical luminescent label.

42. The method of claim 37, wherein the luminescent label is a bioluminescent label or a chemical luminescent label.

43. The method of claim 38, wherein the luminescent label is a bioluminescent label or a chemical luminescent label.

44. The method of claim 39, wherein the luminescent label is a bioluminescent label or a chemical luminescent label.

45. A method for interleaving detection of fluorescent and luminescent signals emitted from a plurality of samples, comprising detecting fluorescent and luminescent signals produced by the plurality of samples, wherein the fluorescence excitation light source is automatically turned off or shuttered during interleaved luminescence imaging.